Automated bioreactor system, system for automatically implementing protocol for decellularizing organ, and waste decontamination system

ABSTRACT

An automated bioreactor system for decellularizing an organ includes a main chamber for containing the organ. The system further includes a reagent chamber containing a liquid phase reagent. A reagent conduit delivers the liquid phase reagent to the main chamber, and a perfusion conduit delivers the reagent from the reagent outlet in the main chamber into the organ. A perfusion pump drives the flow of the reagent. A perfusion pressure sensor detects a pressure of the flowing reagent. A control system controls the perfusion pump to drive the flow of the reagent based on a received input representative of a desired pressure and a received input of the detected pressure. The control system may automatically perform all of the steps of a decellularization protocol based on sensor input. An automated waste decontamination system may also be provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a U.S. National Stage of InternationalApplication No. PCT/US2015/047986 filed on Sep. 1, 2015, which claimsthe benefit of U.S. Provisional Patent Application No. 62/044,647 filedon Sep. 2, 2014, the entire disclosures of all of which are incorporatedherein by reference.

FIELD OF THE INVENTION

The system and methods described herein generally related to abioreactor and its systems, and more specifically, to an automatedbioreactor system for decellularizing an organ, a system forautomatically implementing a protocol for decellularizing an organ, anda waste decontamination system for a bioreactor.

BACKGROUND

While allogeneic lung transplantation is the treatment for end-stagelung disease, the number of patients awaiting lung transplantation issteadily growing and only a small portion of patients receives organtransplantation because of the limited availability of donor organs. Forexample, chronic obstructive pulmonary disease (COPD) affects over 64million people worldwide. The World Health Organization has predictedthat by 2030 COPD will become the third leading cause of mortality. Asanother example, pulmonary arterial hypertension (PAH) affects thevasculature of the lungs and can result in right heart failure anddeath. While there are FDA-approved treatments for PAH, there are nocures, leaving lung transplantation the only option for some patients.Quite simply, there are not enough organs to meet demand.

Even in patients that receive organs available for transplantation,clinical success of lung transplantation in a patient is hampered byimmunosuppression and chronic rejection, which may occur even yearsafter the patient has undergone organ transplant. In a best casescenario, a patient holds at bay these problems by taking medications,which can come with their own serious side effects, for the remainder ofthe patient's life.

Tissue engineering presents an alternative to classic transplantation.This kind of regenerative approach has the potential to effectivelybypass the limitations imposed by tissue donor pools and preventallograft rejection by providing three-dimensional scaffolds for theseeding of autologous or stem cells that are specific to a particularpatient. The technology involves treating an organ with a series ofdetergents, salts, and/or enzymes to completely remove cellular materialin a process known as decellularization, while leaving the extracellularmatrix intact, such that the matrix may serve as scaffold for thesubsequent regeneration of tissue. Decellularization commonly involvessequential perfusion of an organ with a series of detergents andrepetitive washes to remove residual DNA and other cellular debris fromthe organ. This can result in a scaffold in which extracellular matrix(ECM) proteins, organ architecture, and vasculature are retained. Inother words, the scaffold retains the organ's structural features, butis devoid of living cells or cell components. Since cellular antigensthat stimulate immune rejection are commonly found on the cell surface,removal of such antigens may reduce the risk of rejection afterrecellularized scaffolds are implanted into patients. For lungs, thedecellularization process is particularly complex, as it requires thepreservation of airways and alveoli as well as the pulmonary capillarybed to ensure the integrity of the gas exchange tissue.

Human intervention and manipulation during manually performeddecellularization protocols enhance the risk of contamination, decreaseconsistency of the final product, and may adversely affect thethree-dimensional structure and microarchitecture of the resultingscaffold, thus wasting considerable time and resources with apotentially unviable product.

SUMMARY

It is desired to provide an improved bioreactor system and systemsrelated thereto.

One embodiment of the present invention relates to an automatedbioreactor system for decellularizing an organ including a main chamberconfigured to contain the organ and having at least one reagent inlet,at least one reagent outlet, and at least one perfusion inlet. Thesystem further includes at least one reagent chamber containing a liquidphase reagent and at least one reagent conduit configured to deliver theliquid phase reagent from the at least one reagent chamber to the mainchamber through the at least one reagent inlet. At least one perfusionconduit is configured to deliver the liquid phase reagent from the atleast one reagent outlet into the organ through the at least oneperfusion inlet. At least one perfusion pump is configured to drive theflow of the liquid phase reagent through the at least one perfusionconduit. At least one perfusion pressure sensor detects a pressure ofthe liquid phase reagent flowing through the at least one perfusionconduit. A control system receives an input representative of a desiredpressure of the liquid phase reagent flowing through the at least oneperfusion conduit, receives an input of the pressure detected by the atleast one perfusion pressure sensor, and outputs a signal to control theat least one perfusion pump to drive the flow of the liquid phasereagent based on the received input representative of the desiredpressure and the received input of the detected pressure.

Another embodiment of the present invention related to a system forautomatically implementing a protocol for decellularizing an organincluding at least one perfusion pressure sensor configured to detect apressure of liquid flowing into the organ, at least one perfusion valveconfigured to control the flow of liquid into the organ, and at leastone perfusion pump configured to drive the flow of liquid into theorgan. The system further includes a control system configured toreceive a protocol with steps for perfusing the organ, receive inputfrom the at least one perfusion pressure sensor, and control the atleast one perfusion valve and the at least one perfusion pump toautomatically perform all of the steps of the protocol to decellularizethe organ.

Yet another embodiment of the present invention relates to a wastedecontamination system for a bioreactor including a main chamberconfigured to contain an organ for perfusing, a waste chamber configuredto receive waste fluid from the main chamber, and a waste conduitconfigured to deliver the waste fluid from the main chamber to the wastechamber. The system further includes a decontamination fluid chamberconfigured to store decontamination fluid and a decontamination conduitconfigured to introduce the decontamination fluid from thedecontamination fluid chamber into the waste fluid.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic view of a bioreactor for the automated bioreactorsystem according to one exemplary embodiment.

FIG. 2 is a block diagram of a control system for the bioreactor of FIG.1 according to one exemplary embodiment.

FIG. 3 is a screenshot illustrating one embodiment of a main screen of auser interface of the control system of FIG. 2.

DETAILED DESCRIPTION

Overview

The automated bioreactor system described herein provides for abioreactor that can be configured to perform all of the steps of adecellularization protocol automatically, with little to no user inputneeded for successful execution of the protocol after its initiation.The automated bioreactor system includes a main chamber wheredecellularization takes place and a series of reagent chambers thatsupply the main chamber with the reagents needed to facilitatedecellularization in a continuous closed circuit. A control system,having a controller and user interface, is incorporated into theautomated bioreactor system to fully automate the decellularizationprocess in the closed system by controlling the valves and pumps thatdirect the flow of reagents throughout the steps of the protocol. Byproviding for a complete automation of the decellularization process,direct human interface with the components of the bioreactor is reduced,thereby increasing sterility, consistency, and efficiency in theprocess, which improves the likelihood of obtaining a more viable, andstructurally sound decellularized organ for later transplantation.Moreover, the automated bioreactor system allows for a central controlsystem capable of simultaneously performing multiple protocols acrossmultiple bioreactors, further increasing efficiency and consistencyacross multiple systems.

While the processes described below are in the context of adecellularization protocol of a lung, the automated bioreactor systemdescribed herein is not limited to such a protocol or organ. Forexample, the automated bioreactor system may also be utilized to performother applications for which the perfusion of an organ may be required.These applications may include, but are not limited to, processes wheredecellularized lungs are required as well as any downstream analyses ofresulting scaffold material, the recellularization of the scaffold, andthe engineering of a nascent, functional lung. In addition, theautomated bioreactor system may also be utilized to perfuse anyappropriate organ or organ part. Thus, the term “organ” includes wholeorgans as well as any functional portions of the organ, such as a lobeof a lung.

FIG. 1 shows an embodiment of an automated bioreactor system 100 forperforming an entire decellularization protocol of a porcine lung isshown. The automated bioreactor system 100 of FIG. 1 generally includesa main chamber 15 configured to contain a lung 5 to be perfused, achamber fill section 10, and a perfusion section 20. A wastedecontamination system 30 may also be provided for the automatedbioreactor system 100.

As shown in FIG. 2, the automated bioreactor system 100 further includesa controller 50, which is electrically coupled to, and configured tocontrol, various components within each of the sections of thebioreactor, as will be described in more detail below. In addition, auser interface 70 is further connected to and communicates with thecontroller 50 for user management of the automated bioreactor system100, as will be described in more detail below.

Main Chamber

The main chamber 15 may be any appropriate chamber capable of creatingand maintaining a sterile, sealed environment for the perfusion of theorgan or lung 5. The main chamber 15 preferably includes a reagent inlet14, a reagent outlet 23, a perfusion inlet 24, and waste outlets 31, 32,which are described in further detail below. In addition, the mainchamber 15 is positioned on top of a weight sensor 60, which isdescribed in further detail below. In some embodiments, at leastportions of the body of the main chamber 15 are transparent ortranslucent, so a user can view the organ during use of the bioreactorsystem. In some embodiments, the main chamber 15 will have inlets formanipulation of the interior of the main chamber during use. Thismanipulation can occur by physically inserting objects, such as toolsand instruments, into the main chamber 15 or can provide an interfacefor a user to manipulate objects already present in the chamber. Forexample, the inlets can be designed such that the main chamber 15 can beused as a glove box.

Chamber Fill Section

As shown in FIG. 1, the chamber fill section 10 preferably includes aplurality of reagent chambers 11, each fluidly connected to the mainchamber 15 via a reagent conduit, which may take the form of tubing 40.While multiple reagent chambers 11 are depicted, fewer or greaternumbers of chambers may be utilized. The reagent chambers 11 areconfigured to hold a specific type of liquid phase reagent needed for agiven step in the decellularization protocol. The liquid phase reagentcan be in the form of a solution, emulsion, or suspension. The reagentchambers 11 may be any suitable container to hold the reagents, such ascarboys, syringes, or the like. In addition, the reagents contained ineach of the reagent chambers 11 may be any suitable reagent necessary todecellularize an organ, such as phosphate-buffered saline (PBS), sodiumdodecylsulfate (SDS), Triton X-100, or the like. The tubing 40 of thereagent conduit may comprise of any conventional material appropriatefor the transfer of the reagents and biomaterials and for maintainingsterile connections between the chambers, such as silicon tubing or thelike. As shown in FIG. 1, the reagent conduit, e.g., the tubing 40,allows the liquid phase reagents to flow into the main chamber 15through a connection to the reagent inlet 14.

Positioned between the reagent chambers 11 and the main chamber 15 are areagent valve manifold 12 and a reagent pump 13. The reagent valvemanifold 12 controls the flow of a liquid phase reagent from a givenreagent chamber 11 to the main chamber 15, which houses the lung 5. Thereagent valve manifold 12 includes a plurality of valve ports. Thenumber of valve ports provided on the reagent valve manifold 12 may beany appropriate number to allow for an in-line flow of as many liquidphase reagents as needed for the protocol. As shown in FIG. 1, thereagent valve manifold 12 includes five valve ports, allowing for thecontrol of flow of up to five reagent chambers 11 through the tubing 40that connects to the reagent inlet 14 of the main chamber 15. Thereagent valve manifold 13 may also be any appropriate valve mechanismthat can control the flow of a liquid phase reagent from a specificreagent chamber 11 to the chamber fill inlet 14 of the main chamber 15.Preferably, the reagent valve manifold 13 includes reagent valves in theform of solenoid pinch-type valve mechanisms. These solenoid pinch-typevalve mechanisms can control the flow of liquid phase reagent throughthe tubing 40, which is flexible when using the pinch-type valvemechanism, by applying a force to the tubing 40 to impede flow of theliquid phase reagent. Such a valve mechanism is an external cut-offdevice, which minimizes contact with the reagents contained within thetubing 40, thereby increasing sterility of the system.

The reagent pump 13 is connected to the tubing 40 downstream to thereagent valve manifold 12. The reagent pump 13 drives the flow of aliquid phase reagent from a given reagent chamber 11 into the reagentinlet 14. The reagent pump 13 may be any appropriate mechanism for thepumping of fluid into the main chamber 15. In the embodiment shown inFIG. 1, the reagent pump 13 is a double head peristaltic pump to allowfor increased flow rate and pressure, and to maintain sterility of theliquids within the tubing 40. As shown in FIG. 2, both the reagent valvemanifold 12 and the reagent pump 13 are electrically coupled to, andconfigured to be controlled by, the controller 50.

As described above, the main chamber 15 is placed on the weight sensor60, such as, for example, a scale. The weight sensor 60 is configured todetect a weight present in the main chamber 15. As shown in FIG. 2, theweight sensor 60 communicates a detected weight to the controller 50.

Perfusion System

An embodiment of the perfusion system 20 is shown in FIG. 1. It ispreferably configured as a closed circulation system having a perfusionconduit, in the preferred form of additional tubing 40, leading from areagent outlet 23 of the main chamber 15 to a perfusion inlet 24 of themain chamber 15, such that a liquid phase reagent contained in the mainchamber 15 can be re-circulated and perfused through the lung 5. Thetubing 40 forming the perfusion conduit extends further into the mainchamber 15 through the perfusion inlet 24 and into the lung 5 containedin the main chamber 15. As illustrated in FIG. 1, the tubing 40 may beconfigured to extend into the pulmonary artery of the lung 5 such thatperfusion can occur through the cardiopulmonary circuit of the lung 5.

The flow of the liquid phase reagent through the lung 5 is driven by aperfusion pump 21, which, as shown in FIG. 2, is electrically coupledto, and configured to be controlled by, the controller 50. As shown inFIG. 1, the perfusion pump 21 may be a single head peristaltic pump. Apulse dampener 22 may be included and positioned between the perfusionpump 21 and the main chamber 15. The pulse dampener 22 may serve toavoid entry of bubbles into the liquid phase reagent before perfusinginto the lung 5, or may serve to prevent damage to the lung 5 caused bypulses in flow that may be generated by the perfusion pump 21 duringoperation. The pulse dampener 22 may be any appropriate closed containerconfigured to hold residual fluid circulating in the perfusion system.In addition, the pulse dampener 22 may be further provided with a stirbar in order to reduce the chance of stagnation in the well of the pulsedampener 22, a condition which could lead to contamination. Furthermore,the pulse dampener 22 may allow for the reduction of noise in an outputwaveform of pressure in the flow of the liquid phase reagent to the lung5. This, in turn, may allow for the pressure to be measured moreaccurately by a perfusion pressure sensor 25, which is described in moredetail below.

Connected to the tubing 40 upstream from the perfusion inlet 24 of themain chamber 15 is a perfusion pressure sensor 25 for detecting thepressure of the liquid phase reagent flowing through the tubing 40 andinto the lung 5. As shown in FIG. 2, the perfusion pressure sensor 25 isfurther configured to communicate a detected pressure of the liquidphase reagent with the controller 50. The perfusion pressure sensor 25may be configured to detect a pressure of the liquid phase reagent atany point upstream from the perfusion inlet 24. As shown in FIG. 1, theperfusion pressure sensor 25 is connected to the tubing 40 such that theperfusion pressure sensor 25 detects a pressure at the perfusion inlet24. The perfusion pressure sensor 25 may be placed outside the chamberat a height approximately matching the height at which the lung 5 floatsin the main chamber 5 during perfusion.

Waste Decontamination System

As shown in FIG. 1, a preferred embodiment of the waste decontaminationsystem 30 generally includes a decontamination fluid chamber 34 and awaste chamber 35. The waste chamber 35 is fluidly connected to receivewaste fluid from the main chamber 15 via a waste conduit, which may bein the form of tubing 40, while the decontamination fluid chamber 24 isfluidly connected to the waste chamber 35 via a decontamination conduit,which may be formed by additional tubing 40. The decontamination fluidchamber 34 is configured to hold a decontamination fluid, such as, forexample, bleach, while the waste chamber 35 is configured to hold amixture containing the waste fluid from the main chamber 15 and thedecontamination fluid from the decontamination fluid chamber 34.

As shown in FIG. 1, positioned between the main chamber 15 and wastechamber 35 are two drain valves 33, which control the flow of wastefluid from the main chamber 15 through waste outlets 31, 32. Asdescribed above with the valve manifold 12, the drain valves 33 may alsoinclude solenoid pinch valves as the valve-closing mechanism forenhanced sterility of the closed system. In addition, the waste outlets31, 32 may also each be provided with a one-way valve in order toprevent the flow of waste fluid back into the main chamber 15.

Connected downstream from the drain valves 33 is a decontamination pump36, which, as shown in FIG. 2, is electrically coupled to, andcontrolled by, the controller 50. The decontamination pump 36 drives theflow of waste fluid from the main chamber 10 to the waste chamber 27. Inaddition, the decontamination pump 36, as shown in FIG. 1, may alsodrive the flow of decontamination fluid from the decontamination fluidchamber 34 to the waste chamber 35. As illustrated in FIG. 1, thedecontamination pump 55 may take the form of a four-head peristalticpump, in which two pump heads are configured to drive the flow of wastefluid within the tubing 40 that connects the main chamber 15 to thewaste chamber 35, and the other two pump heads are configured to drivethe flow of decontamination fluid from the decontamination fluid chamber34 to the waste chamber 35.

In some instances, certain decontamination protocols require that wastefluid be mixed with a specific proportion of decontamination fluidbefore the waste fluid can be safely and properly drained. For example,before disposal, waste fluid may be mixed with bleach in a desireproportion (e.g., 10%) of the total volume of the waste fluid. Toachieve this, in one embodiment, the decontamination conduit leadingfrom the decontamination fluid chamber 34 to the waste chamber 35 has adiameter that is smaller by the desired proportion (e.g., 1/10 the size)relative to the diameter of the waste conduit leading from the mainchamber 15 to the waste chamber 35. And, as illustrated in FIG. 1,because the decontamination pump 36 drives the flow of both the wastefluid from the main chamber 15 and the flow of decontamination fluidfrom the decontamination fluid chamber 34, the decontamination pump 36may be configured to pump at a constant flow rate such that thedecontamination fluid can be mixed in a proper proportion to the rate offluid flowing into the waste chamber 35. Moreover, as shown in FIG. 1,to ensure adequate mixing of the decontamination fluid with the wastefluid, the decontamination fluid may be introduced into the waste fluidin-line within the waste conduit and upstream from the waste chamber 35.However, the decontamination fluid may also be introduced into the wastefluid directly in the waste chamber 35 and subsequently mixed within thewaste chamber 35.

In addition, separate pumps may be alternatively utilized to drive theflow of waste fluid from the main chamber 15 to the waste chamber 35 andthe flow of decontamination fluid chamber 34 to the waste chamber 35. Inthis sense, the controller 50 may then control each of thedecontamination pump 36 and the additional pump to drive the flow ofwaste fluid from the main chamber 10 and the decontamination fluid fromthe decontamination fluid chamber 34, respectively. Each of the pumpsmay be controlled to operate at a respective flow rate such that thedecontamination fluid mixes with the waste fluid in a proportionnecessary for safe and proper disposal.

As shown in FIG. 1, a drain conduit, in the form of additional tubing40, extends from the waste chamber 35 to a drain 37. Positioned betweenthe waste chamber 35 and the drain 37 is a drain pump 38, which drivesthe flow of waste fluid and decontamination fluid mixture from the wastechamber 35 to the drain 37. A shown in FIG. 2, the drain pump 38, whichmay be a double head peristaltic pump, is electrically connected to, andcontrolled by, the controller 50. Alternatively, however, the drain pump38 may be omitted and the drain conduit may instead extend from thebottom of the waste chamber 35. A valve, such as a solenoid pinch valve,may be included down stream of an outlet of the waste chamber 35. Thecontroller 50 may then be configured to control the valve such that,when opened, the waste fluid and decontamination fluid mixture isallowed to flow to the drain 37 via gravity.

As further shown in FIG. 1, a plurality of secondary containmentchambers 39 are shown, which serve as a secondary measure to preventwaste fluid from flowing outside the automated bioreactor system 100. Asillustrated in FIG. 1, the main chamber 15, the decontamination fluidchamber 34, and the waste chamber 35 are each placed in a secondarycontainment chamber 39. The secondary containment chambers 39 serve tohold any overflow of fluids from each of these chambers. As shown inFIG. 1, each of the secondary containment chambers 39 is connected to aplurality of drain conduits, each formed by tubing 40, which allows anyfluids contained in the secondary containment chambers 39 to flow intothe drain 37.

The automated bioreactor system 100 is not limited to the sensors shownin FIG. 1, and may incorporate the use of any additional sensors thatmay measure a variable relevant to the perfusion process. For example, atemperature control system may be included. The temperature controlsystem may have temperature sensors placed adjacent to the main chamber15 in order to detect a temperature of the liquid phase reagents andlung 5 contained in the main chamber 15. The temperature control systemmay further include a heating and/or cooling element configured to heatand/or cool the temperature of the liquid phase reagents and lung 5contained in the main chamber 15. The temperature sensors may then beconfigured to communicate the detected temperature to the controller 50.The controller 50 may then be configured to control the heating and/orcooling element in order to adjust the temperature of the liquid phasereagents and lung 5 to a safe temperature for the decellularizationprocess. By controlling for temperature within the main chamber 15,sterility of the system and the structural integrity of the organ may befurther maintained. Additional sensors may also include, but are notlimited to, pH sensors, conductivity sensors, and the like.

As described above, the automated bioreactor system 100 includes abioreactor for the decellularization of a lung in a closed, sterileenvironment. By limiting human interaction with the mechanisms thatcontrol the decellularization process and by minimizing contact of thesystem's moving parts (e.g., valve mechanism, pumps) with the internalcomponents (e.g., liquid reagents, organ), a more sterile bioreactorsystem may be achieved, thereby increasing the likelihood of a viableand structurally sound scaffold after the decellularization process.

Control Process

An overview of the control process for automating an entire protocol fordecellularizing a lung 5 will now be described with continuing referenceto FIGS. 1 and 2. In general, the control system may automaticallyexecute all of the steps of an entire protocol by performing,sequentially and/or simultaneously, three types of processes: a chamberfill process, a perfusion process, and a waste decontamination process.

The protocol can be any of the known protocols for perfusing ordecellularizing an organ, as well as being a new protocol. The protocolmay be provided to the system in any of a variety of ways. For example,the protocol may be uploaded as part of a data spreadsheet.Alternatively, the protocol, or a number of protocols, may be stored onthe system for selection by the user. As a further alternative, thestored protocols may be modified by the user.

The control system, or controller 50 thereof, can be a general purposeprocessor, a digital signal processor (DSP), an application specificintegrated circuit (ASIC), a field programmable gate array (FPGA) orother programmable logic device, discrete gate or transistor logic,discrete hardware components, or any combination thereof designed toperform the functions described herein. A general-purpose processor maybe a microprocessor, but in the alternative, the processor may be anyconventional processor, controller, microcontroller, or state machine. Aprocessor may also be implemented as a combination of computing devices,e.g., a combination of a DSP and a microprocessor, a plurality ofmicroprocessors, one or more microprocessors in conjunction with a DSPcore, or any other such configuration. The steps of a method oralgorithm described in connection with the embodiments disclosed hereinmay be embodied directly in hardware, in a software module executed by aprocessor, or in a combination of the two. A software module may residein RAM memory, flash memory, ROM memory, EPROM memory, EEPROM memory,registers, hard disk, a removable disk, a CD-ROM, or any other form ofstorage medium known in the art. An exemplary storage medium is coupledto the processor such that the processor can read information from, andwrite information to, the storage medium. In the alternative, thestorage medium may be integral to the processor. The processor and thestorage medium may reside in an ASIC. The ASIC may reside in a userterminal. In the alternative, the processor and the storage medium mayreside as discrete components in a user terminal. In one or moreexemplary embodiments, the functions described may be implemented inhardware, software, firmware, or any combination thereof. Such hardware,software, firmware, or any combination thereof may be part of orimplemented with any one or combination of the servers, databases,associated components in the system, components thereof, and/or thelike. If implemented in software, the functions may be stored on ortransmitted over as one or more instructions or code on acomputer-readable medium. Computer-readable media includes both computerstorage media and communication media including any medium thatfacilitates transfer of a computer program from one place to another. Astorage media may be any available media that can be accessed by acomputer. By way of example, and not limitation, such computer-readablemedia can comprise RAM, ROM, EEPROM, CD-ROM or other optical diskstorage, magnetic disk storage or other magnetic storage devices, or anyother medium that can be used to carry or store desired program code inthe form of instructions or data structures and that can be accessed bya computer. In addition, any connection is properly termed acomputer-readable medium. For example, if the software is transmittedfrom a website, server, or other remote source using a coaxial cable,fiber optic cable, twisted pair, digital subscriber line (DSL), orwireless technologies such as infrared, radio, and microwave, then thecoaxial cable, fiber optic cable, twisted pair, DSL, or wirelesstechnologies such as infrared, radio, and microwave are included in thedefinition of medium. Disk and disc, as used herein, includes compactdisc (CD), laser disc, optical disc, digital versatile disc (DVD),floppy disk, and Blu-Ray disc where disks usually reproduce datamagnetically, while discs reproduce data optically with lasers.Combinations of the above should also be included within the scope ofcomputer-readable media.

A chamber fill process begins with the controller 50 outputting a signalto the reagent valve manifold 12 to open one or more of the valves(while, at the same time, closing any unnecessary valves for theprocess) to allow for the flow of the liquid phase reagent contained inthe given reagent chamber 11. At the same time, the controller 50 alsooutputs a signal to the reagent pump 13 to drive the flow of the liquidphase reagent(s) from the reagent chamber(s) 11 connected to thenow-opened valve line(s) into the tubing 40 leading to the reagent inlet14. The reagent pump 13 may be driven according to the protocol and/oruser settings as specified. For example, the controller 50 may output tothe reagent pump 13 to pump at a constant flow rate (e.g., 100 ml/min)or at a constant pressure (e.g., 30 mmHg). If the reagent pump 13 is aperistaltic pump, the controller 50 may further output to the reagentpump 13 operate in a given direction (i.e., clockwise orcounterclockwise).

As the main chamber 15 is being filled with the desired liquid phasereagent(s), the controller 50 continuously receives an input of a weightof the main chamber 15 as detected by the weight sensor 60, as shown inFIG. 2. Using this detected weight, the controller 50 then determines avolume of liquid phase reagent present in the main chamber 15 at thatgiven time. For example, the controller 50 may determine the volume ofliquid phase reagent based on both the detected weight of the mainchamber 15 and a known density of the liquid phase reagent being used,which may be pre-stored in the program before execution of the protocol.By continuously monitoring the weight of the main chamber 15, and thus,the volume of liquid phase reagent present in the main chamber 15, thecontroller 50 may then determine when the volume of liquid phase reagentrequired for the chamber fill step of the protocol has been reached.

In addition, the controller 50 may also be configured to determine ifand/or when a maximum volume of liquid reagent has been introduced intothe main chamber 15. In this case, the controller 50 is programmed,before the execution of the protocol, with a pre-stored threshold volumethat may represent the maximum volume of fluid that the main chamber 15is capable of holding or, alternatively, some proportion of the maximumvolume. During execution of a chamber fill step, if the controller 50determines that the main chamber 10 has been filled such that thethreshold volume has been reached, the controller 50 may be configuredto stop the continuation of the chamber fill step (by stopping operationof the reagent pump 13) even if the chamber fill step still requiresadditional liquid reagent to be introduced into the main chamber 15. Thecontroller 50 may then automatically proceed to the next step in theprotocol (e.g., a perfusion step or a drain step). In this sense, bycontinuously monitoring the weight of the main chamber 10, thecontroller 50 may prevent the overfilling of the main chamber 15,thereby providing a safeguard for the system.

During a perfusion process, with all valves closed in the system, thecontroller 50 outputs a signal to control the perfusion pump 21 in orderto drive the flow of the liquid phase reagent contained in the mainchamber 15 into the lung 5 (e.g., into the pulmonary artery of the lung5). As similarly described above with the chamber fill process, thecontroller 50 may output a signal to control the perfusion pump 21 inorder to drive the flow of the liquid phase reagent at a constantpressure or a constant flow rate, as defined by the protocol. Inaddition, if a peristaltic pump is utilized, the controller 50 maydirect the perfusion pump 21 to operate in a given direction as well.

During perfusion, the controller 50 continuously receives an input of apressure of the flow of the liquid phase reagent to the lung 5 detectedby the perfusion pressure sensor 25, as shown in FIG. 2. The controller50 may be also configured to receive an input of a desired pressureand/or flow rate of the liquid phase reagent into the lung 5. Thedesired pressure and/or flow rate may be defined by the protocol or maybe user-specified and may be inputted into the control system by aninput representative of the desired pressure through the perfusionconduit. Based on the received detected pressure and the received inputrepresentative of the desired pressure, the controller 50 may thenoutput a signal to control the perfusion pump 21 such that the desiredpressure and/or flow rate into the lung 5 is continuously maintained.The controller 50 may also be configured to detect if and/or when thedetected pressure has deviated by a pre-stored threshold amount from thedesired pressure and/or flow rate of the liquid phase reagent into thelung 5. For example, the controller 50 may be configured to detect ifand/or when the detected pressure is 15% higher or lower than theprotocol-defined pressure and/or flow rate. The controller 50 may thenbe further configured to communicate to the user interface 70 thisdeviation, which may subsequently alert the user of the deviation fortroubleshooting purposes.

After completion of a perfusion step, the controller 50 may beconfigured to initiate additional wash steps using, for example, wateror saline, to rinse away residual liquid reagents from the lung 5 or theperfusion conduit before proceeding to the next step. In this case, thecontroller 50 may output a signal to the valve manifold 14 of thechamber fill section 10 to open the valve and allow flow from a reagentchamber 11 configured to hold a washing fluid. The controller 50 maythen control the reagent pump 13 and the perfusion pump 21 to drive theflow of the washing fluid through the system.

During a waste decontamination process, the controller 50 outputs asignal to open the drain valves 33 to allow the flow of waste fluid fromthe main chamber 15 to the waste chamber 35. Simultaneously, thecontroller 50 outputs a signal to the decontamination pump 36 to directthe flow of the waste fluid from the main chamber 15 to the wastechamber 35. As described above and shown in FIG. 1, the decontaminationpump 36 may be also configured to direct the flow of decontaminationfluid from the decontamination fluid chamber 34 to the waste chamber 35.The controller 50 may then output a signal to the decontamination pump36 to direct flow of both the waste fluid and the decontamination fluidthrough the tubing 40 such that the two fluids mix in the properproportions. Alternatively, if two separate pumps are used to direct theflow of the waste fluid and the decontamination fluid, respectively, thecontroller 50 may output a signal to each of the pumps to direct a flowof the respective fluid at a rate that will ensure proper mixing of thetwo fluids. As similarly described above, the controller 50 may output asignal to the decontamination pump 36 to pump the waste fluid and/ordecontamination fluid flowing through the tubing 40 at a constantpressure or a constant flow rate.

As the waste decontamination process proceeds, the controller 50continuously monitors the volume of waste fluid drained from the mainchamber 15 by receiving input of weight detected by the weight sensor60. Once the required volume of waste fluid has been drained from themain chamber 15, as defined by the protocol, the controller 50 outputs asignal to the valves 33 to close. Once the waste fluid anddecontamination fluid mixture has mixed and settled in the waste chamber35 for the required amount of time to ensure adequate decontamination,the controller 50 then outputs a signal to the drain pump 38 to drivethe flow of the fluid contained in the waste chamber 35 into the drain37 for disposal. Alternatively, as described above, the controller 50may be configured to output a signal to open a valve located at thebottom of the waste chamber such that the fluid contained in the wastechamber 35 is drained via gravity.

As mentioned above, and as shown in FIG. 2, the controller 50communicates with a user interface 70. The user interface 70 allows auser to remotely write a decellularization protocol for uploading to thecontrol system, and then set-up and manage the automated protocolexecuted by the automated bioreactor system 100. An embodiment of a mainscreen of the user interface 70 is shown in FIG. 3 and described infurther detail below.

The user interface 70 allows the user to define a series of steps (a“recipe”) that contains the operating information for the pump and valvefunctions. The user-defined recipe serves to guide the controller 50 asto the appropriate components to control at a given time duringexecution of the entire protocol. The series of steps may be written bythe user using any appropriate text-based program (remotely or withinthe user interface 70), which may then be uploaded into the controlsystem for execution. In addition, the user-defined series of steps maybe stored in the control system for later recall and execution, therebyallowing for an easy selection and execution of commonly used protocolsand set-ups.

Before beginning the automatic decellularization process, the userinterface 70 allows the user to set system settings of the givenbioreactor being used for the decellularization protocol under a systemsettings command 89, which may bring up a separate window of the userinterface 70 to allow the user to establish settings for the bioreactorsystem. Here, pressure inputs, digital outputs for the solenoid rack,and pump task may be configured here to ensure that data is readproperly from the devices. Once the desired system settings have beenselected by the user, the user may then begin execution of the selectedprotocol by the automated bioreactor system 100 through the userinterface 70 via a start command. A reinitialize hardware command 90 mayalso be included in order to initialize the hardware before starting anew recipe.

During execution of the automatic decellularization process, the userinterface 70 allows the user to remotely monitor and manage theexecution of the protocol. For instance, under a section A of the mainscreen of the user interface 70 as shown in FIG. 3, the user may start,stop, or pause the execution of the protocol as it progresses usingcommand buttons 71. While pausing the execution using the command button71 temporarily stops all pumps, the user can still turn on the pumps byusing the override parameters contained in section E, which is describedin further detail below. In addition, the command buttons 71 may furtherincludes a skip command, allowing the user to skip a current step in theprotocol and proceed to a subsequent step.

The user interface 70 may also provide the user with real-timeinformation regarding the status of the bioreactor as the protocol isexecuted. For example, as shown in FIG. 3 under section C, the userinterface 70 may allow the user to view the time remaining in theexecution of the current step and/or overall protocol under indicators73. The user interface 70 may also provide a viewable list of all thesteps of the protocol under a tab 72 and indicate the current step beingexecuted by the system under an indicator 74. In addition, the userinterface 70 may allow the user to monitor the input received by thecontroller 50 from the various sensors present in the system. Forexample, the user interface 70 may be configured to display a waveformchart 75 showing pressure as detected by the pressure sensor 25 overtime under section D of the main screen shown in FIG. 3. Aspects of thewaveform chart may be manipulated by the user with appropriate graphicalmanipulation tools (e.g., zoom, scale, drag, etc.). Actual, measuredvalues of the pressure and flow rate determined by the controller 50based on the detected pressure input received from the pressure sensor25 may also be displayed via a table 76. Moreover, for systems utilizingmultiple pressure sensors, a toggle button 87 may be provided such thatthe user can switch and/or add real-time information regarding thepressures detected by each of the pressure sensors. Similarly, thevolume of liquid phase reagent contained in the main chamber 15 asdetermined by the controller 50 may also be displayed in real-timethrough an indicator 77 under section D. The user may zero an/orcalibrate any pressure sensors utilizing a recalibrate command 88.However, preferably the recalibrate command 88 is only active before arecipe is started and becomes inactive while an automateddecellularization protocol is active.

In addition, the user interface 70 may also be configured to inform theuser of the current operating states of the valves and pumps. Forexample, the user interface 70 may indicate to the user the currentoperating status of a given pump (e.g., idle, testing, alarm state)under a tab 78. Under this tab, errors for the pressure inputs, thepumps, and the solenoids may be displayed. Scale debug values may alsobe displayed, which include measurements for volume display and are usedto calculate the volumes for the fill steps.

The overall main status of the system (e.g., idle, testing/running,alarm state) may be indicated to the user through an indicator 79 undersection C of the main screen of the user interface 70. As describedabove, the controller 50 may be configured to detect if a detectedpressure of a flow rate driven by a given pump deviates by more than apredetermined threshold. If such deviation occurs, the controller 50 maythen communicate this status to the user interface 70, which may beconfigured to alert the user of the alarm state of the pump (e.g.,indicator 79 changes from green to yellow when the pressure readingdeviates by 15% from the pressure set point of at least one pump, thenfrom yellow to red when the pressure reading deviates by more than 30%from the pressure set point of at least one pump).

To allow for troubleshooting, the user interface 70 may contain overrideparameters under section E of the main screen as shown in FIG. 3. Inthis section, the user can alter pump settings and valve status duringexecution of the protocol in order to deviate from the uploadedprotocol. For example, pump settings may be overridden via a pumpcontrol section 82. Using the pump control section 82, the user may turna pump on or off, change the mode of operation of the pump (e.g.,constant flow rate to constant pressure), change the direction ofoperation of the pump (e.g., clockwise to counterclockwise), and/orchange the flow rate and/or pressure set point. Similarly, the valvesettings may also be changed under a valve control section 83, in whichthe user may open or close a specific valve in the system. Fluidsettings may also be manually changed via a fluid control section 84. Asshown in FIG. 3, the user may select a fluid from a bank of drop-downmenus, which correspond to the fluid flow controlled by the valves shownin valve control section 83. Alternatively, the user may choose tomanually insert a specific fluid not contained in the drop-down menuand, thus, may utilize a fluid settings command 85 to update the system.Once selections are updated, the density information may then be passedto the control system for calculation of the display volume and targetfill volume of the main chamber 15. A vessel settings command may alsobe included to allow the user to enter information about the bioreactorthemselves. Thus, the user interface 70 allows for continuous monitoringand management of the automatic decellularization process in order toallow for a more effective and efficient control of the overallprotocol.

The user interface 70 may also include a notification settings command81 that allows the user to remotely receive notifications regarding thesystem as it executes the protocol. This settings command 81 may allowfor automatic notification, in the form of, for example, email, of anychanges in a given step by the control system and periodic updates onthe state of the system as a whole. In addition, the user interface 70may allow for the insertion of time-stamped comments to the data log viaa comment command 86, which is saved in the system for later viewing.This may allow the user to note specific changes made to the protocolfor analysis purposes, such as inserting indicating notes about stepstaken during the set-up that might correspond to abnormalities in thedata log, for example, by adjusting a pressure sensor.

The user interface 70 may also allow for the monitoring and managementof the execution of a plurality of protocols, each being executed withina separate bioreactor, such as the one shown in FIG. 1. In this case,the controller 50 acts as a central control system and controls thecomponents of each of the bioreactors according to each of theuser-defined series of steps. The user interface 70 may allow the userto toggle between executed protocols in order to monitor and manage eachseparate protocol simultaneously. By allowing for such a centralizedsystem, several decellularization protocols may be performedsimultaneously, thereby increasing efficiency of the entire process.

Exemplary Protocol for the Preparation of Lung Tissue to beDecellularized by a Bioreactor System

A clean work area is prepared in a biological safety cabinet (BSC). Allsteps of the preparation procedure are performed using good aseptictechnique to minimize introduction of environmental microbialcontaminants. Sterile sleeves and gloves and exchange gloves are used asneeded.

The pluck (i.e., the heart-lung circuit) is unpacked from the shippingcontainer and critically examined for trauma to the vasculature, airway,pleura, and parenchyma. The pluck can then be appropriately dissectedunder sterile conditions. The packaging can be sterilized. The pulmonaryartery cannula is inserted into the pulmonary artery and secured, andthe pluck is dissected such that the heart muscle is removed. Anyremaining heart tissue may be trimmed away. The lung may then beperfused via the pulmonary veins with heparinized saline foranticoagulation during subsequent perfusion in the bioreactor system.The prepared lung is then weighed and transferred to a laminar flow hoodarea for loading into the main chamber when required.

Exemplary Setup of the Bioreactor in the Laminar Flow Hood

With the laminar flow hood blower on, the laminar flow hood is sprayeddown with the appropriate cleaning solution. The autoclaved main chamberis brought near the laminar flow hood, and the outer layer of autoclavewrap is unwrapped. The main chamber is slid into the hood using a ramp.With sleeves and surgical gloves, the inner layer of the autoclave wrapon the main chamber is unwrapped and removed from the hood. Spacersbetween the bioreactor glass and the base are removed. Once all spacersare removed, it is verified that the glass is sitting on the bottomgasket with an even margin around the base.

The autoclave wrap that covers the glass is removed. A person outsidethe laminar flow hood removes the outer layer of the top ring, andbrings it into the hood using sterile gloves. A sterile drape is broughtinto the hood. The top ring is then secured to the metal standoffs usingbolts and tools.

The main chamber lid is brought into the laminar flow hood. The lid isplaced on top of the main chamber, and fasteners attach the lid to thetop ring. Now that the main chamber is closed to the environment, all ofthe ports on the lid and the base are tightened. Using the biowelder,the reagent conduit line is welded to the reagent tubing assembly. Theassembly is welded to a reagent chamber. Using a peristaltic pump, themain chamber is filled.

After filling to the level of the tube drain, the reagent pump isstopped. The perfusion conduit is welded from the base to one end of thepulse dampener. The other side of the pulse dampener is welded to theperfusion conduit at the top of the bioreactor, completing the perfusioncircuit.

Perfusion is started by perfusing solution through the perfusionconduit, and air is removed from the lines using a sterile syringe. Themain chamber is filled to approximately 20 L.

Exemplary Protocol for the Aseptic Installation of the Lung in theLaminar Flow Hood

The perfusion pump is run slowly (25 mL/min) and all air is purged inthe perfusion conduit. The organ (lung) is brought to the laminar flowhood in a draped sterile pan. The cranks on the main chamber lid areloosened. With sterile gloves, the lung is moved into the main chamberand the connection(s) are made.

The perfusion conduit is assessed for any air bubbles. Using the reagentpump, solution is pumped backwards into the 1 x PBS reagent chamberuntil the main chamber is at about 20 L. The reagent conduit is sealedand disconnected from the 1 x PBS reagent chamber. The main chamber isthen removed from the laminar flow hood and transferred to thedecellularization station. The laminar flow hood is disinfected afterinstallation of the lung with the appropriate disinfectant.

Exemplary Automated Lung Decellularization Protocol

Once the main chamber is placed atop the scale, a biowelder is used toweld the following lines and complete the bioreactor fluid circuit: (1)reagent conduit; (2) drain conduit; (3) reagent chamber connections tothe valve manifold.

Perfusion is then started manually at a low rate. About 1-2 L of PBS ismanually filled into the main chamber at 1000 mL/min speed.

The perfusion pressure sensor is placed outside the chamber at a heightof 40 L, matching the height at which the lung will be floating for themajority of the experiment. The perfusion pressure sensor is zeroed bypressing the “Recalibrate” button, then pressure transducer 1 (orwhichever transducer is controlling the perfusion pump, specified by therecipe) is zeroed.

The experiment is initiated by pressing the “Start” button, followingthe prompts for the appropriate recipe file, data save location, andsolution/chamber settings.

A sample automated decellularization process to which a lung set mightbe exposed is as follows: (1) perfuse PBS at 30 mmHG for 2 hours oncerecipe is initiated drain 20 L from the main chamber; (2) perform fivecycles of chamber fill, perfusion, and drain over five days of 0.5% SDS;(3) perform one cycle of chamber fill and drain and one day of exposureof DiH₂O; (4) perform one cycle of chamber fill, perfusion, and drainover one day of 0.5% Triton-X; and (5) perform five cycles of chamberfill, perfusion, and drain over five days of PBS of final wash steps.

Frequent monitoring of the tissue state and bioreactor function isimportant to this process; manual intervention for overflow wastedraining may be required depending on tissue perfusion efficiency.Sampling of chamber fluids may be undertaken during the process forsterility testing or analytics. These samples are taken through theswabbable port on the top of the main chamber.

Exemplary Storage of the Decellularized Tissue

At the end of the experiment, the decellularized tissue will either bestored, sampled and formalin-fixed, or both.

The experiment is stopped on the software interface by pressing the“stop” button. The main chamber is drained manually (drain waste first,if necessary) until no liquid remains in the main chamber. Using aclipster tool or a heat seal tool, all connections from the main chamberto the peripheral tubing are closed off.

Prior to putting the main chamber into the laminar flow hood, theexterior of the main chamber is disinfected. The main chamber is loadedinto the laminar flow hood, then the sterile pan/drape. The lid isremoved. Using sterile gloves, the lung is disconnected from theperfusion conduit and brought into the pan. The pan is removed from thelaminar flow hood and brought into a biosafety cabinet for storage orsampling. The main chamber is removed from the laminar flow hood, andthe unit is disinfected according to protocol.

Exemplary Protocol for Chamber and Station Cleaning

All tubing and fittings downstream of the chamber (drain conduit, waste)should be disposed of in a biohazard waste container. Perfusion conduittubing should be disposed of in this manner as well; however thepressure sensors can be cleaned and saved.

Due to the coating on the main chamber and relative fragility of itscomponents, main chamber parts must be washed by hand with a milddetergent. Clean main chamber parts are allowed to dry.

The waste chamber is stripped of its external tubing—this tubing isdisposed of as biohazard waste. The carboys themselves can be washed inthe parts washer. The cap/dip tube in the waste drain carboys should bewashed by hand. The overfill protection containers that house the wastedrain carboys can be washed by hand if there was a spill during thedecellularization experiment. The bleach carboys need not be washed justfilled up again with bleach when appropriate. The bleach line tubingcircuit should be replaced each decellularization to avoid cracking ofthe fittings that are exposed to bleach.

The residual fluid in the pulse dampener should be decontaminated withbleach and poured down the drain. The pulse dampener itself may bedisassembled and washed. The stir bar may be washed by hand or in thebasket in the parts washer.

Exemplary Operation of the Bioreactor Program

The bioreactor program is opened from the computer desktop and systemsettings are updated if a hardware or software change has occurred.Under System Settings, the following four properties must be set for anormal decellularization process in order to ensure that data will beread properly from the devices: (1) analog input device (pressureinputs); (2) DigOut device (solenoids); (3) analog output task (pumpcontrol); and (4) scale COM port (scale-port connection). Flow ratelimits, which determine the minimum and maximum flow rate in theperfusion line when the perfusion pump is pressure-controlled are alsoset. The PC ID is used as an identifier to which system parameters aresaved. For example, when the PC ID is set to “bioreactor 1”, all theassociated settings are saved to this ID. The PC ID (and associatedsystem settings) that was last used in execution of the program willappear by default upon subsequent executions of the program. The PIDparameters default to a value of 0.1 for Kp, Ki, and Kd. The medianfilter rank is a filter which averages the data points read from thepressure sensor to minimize noise. The larger the value specified, thelarger the observed filter effect. It defaults to a value of 2, but canbe modified to reduce the noise in the pressure chart. A value of 25 hasbeen found to suitable for the automated decellularization process.

The “Email Setting” button will open the Email Configuration screen.Email settings can be changed at any time. A box is checked to receiveemail messages, which include notification of system status changes andperiodic test status emails. The interval at which emails will bereceived is entered. For example, specifying an interval of 01:00:00will send periodic emails every hour. Periodic emails are sent as averification of the current status of the system, steps, pumps, andsolenoid valves. Additionally, system status change emails will be sentwhenever the system status changes, whether by automatic or manualoperation. After all information is appropriately inputted, a test emailis sent by clicking a “Test” button. If the settings are satisfactoryand the test email has been received, the “Save and Exit” button ispressed to save changes and return to the main screen.

By clicking the “Start” button, a Select Recipe box is brought up. Themost recent recipe text file is opened by clicking on the “Open .XLSFile” button. Recipe files are generally located on a desktop folder.The recipe is reviewed to ensure that the proper file was selected, anda “Continue” button is clicked which prompts the user to enter datalogging information. A folder is chosen in which to save the data. Thefolder chosen should reflect the bioreactor station used (4A, 4C, 5A,5C). Data point save frequency (60 seconds) is selected. The “Continue”button is selected once all settings have been entered to start theprocess.

Upon selecting the recipe, the user will be asked to validate that thefluid and vessel settings as appropriate. After selecting the recipe,the user will be prompted one final time before starting the automateddecellularization process. Once started, the recipe can be paused byclicking the ‘Pause’ button on the front panel. A user may need to pauseexecution of the recipe for troubleshooting purposes. In a paused state,the program will display a flashing “Resume” button, which prompts theuser to continue the execution after manual troubleshooting. In a pausedstate, a user may toggle valves, turn pumps on and off, and adjust pumpsettings. Though pumps may be running during a paused state, thebackground color will remain gray, indicative of an idle state. Datacontinues to be recorded even in a paused state. The recipe resumes inthe same step and with the same remaining step time at which it waspaused. If any pump settings were changed during pause, these changeswill persist upon resuming the step. Pumps and the solenoid valves canbe manually set while the recipe is running. A user may need to manuallyadjust these settings for troubleshooting purposes, such as switchingthe perfusion pump to constant pressure instead of constant flow. Theoverride settings will persist for the remainder of the step, as well asthrough a pause and resume action.

Once the recipe has completed the final step, the main status willdisplay “Test Completed”. Data is logged to a file, which can beaccessed via Microsoft Excel. Pressure vs. Timestamp data for theduration of the execution is stored.

Although the foregoing refers to particular preferred embodiments, itwill be understood that the present invention is not so limited. It willoccur to those of ordinary skill in the art that various modificationsmay be made to the disclosed embodiments and that such modifications areintended to be within the scope of the present invention.

Unless otherwise specified, “a” or “an” means “one or more.”

All of the publications, patent applications and patents cited in thisspecification are incorporated herein by reference in their entirety.

What is claimed is:
 1. An automated bioreactor system fordecellularizing an organ, comprising: a main chamber configured tocontain the organ, the main chamber having at least one reagent inlet,at least one reagent outlet, and at least one perfusion inlet; at leastone reagent chamber containing a liquid phase reagent; at least onereagent conduit configured to deliver the liquid phase reagent from theat least one reagent chamber to the main chamber through the at leastone reagent inlet; at least one reagent valve configured to control aflow of the liquid phase reagent through the at least one reagentconduit; at least one perfusion conduit configured to deliver the liquidphase reagent from the at least one reagent outlet into the organcontained in the main chamber through the at least one perfusion inlet;at least one perfusion pump configured to drive the flow of the liquidphase reagent through the at least one perfusion conduit; at least oneperfusion pressure sensor configured to detect a pressure of the liquidphase reagent flowing through the at least one perfusion conduit; aweight sensor configured to detect a weight contained in the mainchamber; and a control system configured to: initiate a chamber fillprocess comprising outputting a signal to open a valve associated withthe reagent conduit to fill the main chamber with a predetermined amountof liquid phase reagent; receive an input of the weight detected by theweight sensor; output a signal to control the at least one reagent valvebased on the detected weight; receive an input representative of adesired pressure of the liquid phase reagent flowing through the atleast one perfusion conduit during a perfusion process; after thechamber fill process is complete, initiate the perfusion processcomprising: receiving an input of the pressure detected by the at leastone perfusion pressure sensor; and outputting a signal to control the atleast one perfusion pump to drive the flow of the liquid phase reagentbased on the received input representative of the desired pressure andthe received input of the detected pressure.
 2. The automated bioreactorsystem of claim 1, further comprising: at least one reagent pumpconfigured to drive the flow of the liquid phase reagent through the atleast one reagent conduit.
 3. The automated bioreactor system of claim2, wherein the control system calculates a volume of the liquid phasereagent contained in the main chamber based on the detected weight and adensity of the liquid phase reagent.
 4. The automated bioreactor systemof claim 1, wherein the input representative of the desired pressure isat least one of a pressure and/or a flow rate.
 5. The automatedbioreactor system of claim 1, wherein the at least one perfusionpressure sensor is configured to detect a pressure at the at least oneperfusion inlet.
 6. The automated bioreactor system of claim 1, furthercomprising: at least one reagent valve configured to control a flow ofthe liquid phase reagent through the at least one reagent conduit,wherein the reagent conduit is flexible and the at least one reagentvalve is a pinch valve that applies a force to the flexible reagentconduit to impede flow of the liquid phase reagent.
 7. The automatedbioreactor system of claim 1, further comprising: at least onetemperature sensor configured to detect a temperature within the mainchamber; and at least one temperature control system configured toadjust the temperature within the main chamber, wherein the controlsystem is further configured to: receive an input of the temperaturedetected by the at least one temperature sensor; and output a signal tocontrol the temperature control system to adjust the temperature in themain chamber.
 8. An automated bioreactor system for decellularizing anorgan, comprising: a main chamber configured to contain the organ, themain chamber having at least one reagent inlet, at least one reagentoutlet, and at least one perfusion inlet; at least one reagent chambercontaining a liquid phase reagent; at least one reagent conduitconfigured to deliver the liquid phase reagent from the at least onereagent chamber to the main chamber through the at least one reagentinlet; at least one temperature sensor configured to detect atemperature within the main chamber; at least one temperature controlsystem configured to adjust the temperature within the main chamber; atleast one perfusion conduit configured to deliver the liquid phasereagent from the at least one reagent outlet into the organ contained inthe main chamber through the at least one perfusion inlet; at least oneperfusion pump configured to drive the flow of the liquid phase reagentthrough the at least one perfusion conduit; at least one perfusionpressure sensor configured to detect a pressure of the liquid phasereagent flowing through the at least one perfusion conduit; and acontrol system configured to: initiate a chamber fill process comprisingoutputting a signal to open a valve associated with the reagent conduitto fill the main chamber with a predetermined amount of liquid phasereagent; receive an input representative of a desired pressure of theliquid phase reagent flowing through the at least one perfusion conduitduring a perfusion process; and after the chamber fill process iscomplete, initiate the perfusion process comprising: receiving an inputof the pressure detected by the at least one perfusion pressure sensor;outputting a signal to control the at least one perfusion pump to drivethe flow of the liquid phase reagent based on the received inputrepresentative of the desired pressure and the received input of thedetected pressure; receiving an input of the temperature detected by theat least one temperature sensor; and outputting a signal to control thetemperature control system to adjust the temperature in the mainchamber.